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Showing 1 to 16 of 16 for “"expressed protein ligation"”.
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Site-Specific Incorporation of Biochemical and Biophysical Probes into Proteins Using Expressed Protein Ligation
<p>Protein engineering can be made far more powerful if a protein is not only expressed recombinantly but also altered covalently using synthetic chemistry. These two methods are brought together in the protein semi-synthesis technique Expressed Protein Ligation (EPL). In EPL, recombinant and …
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Synthesis of Selenium-Alkylated Selenocysteine Derivatives and Their Use in Native Chemical Ligation, Expressed Protein Ligation and the Synthesis of Dehydropeptides
Finally, an improved, scalable synthesis of Fmoc-phenylselenocysteine is reported. The amino acid was coupled to a model peptide on the solid phase. It was demonstrated that the selenide could be oxidatively eliminated to give a dehydropeptide while still bound to the solid resin. In addition, the …
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Induction of Innate and Adaptive Immunity In Vivo by Selective Antibody Targeting of Poly dA:dT and Antigen to Dendritic Cells
… in vivo. Here we illustrate a novel method using expressed protein ligation (EPL) to attach the immune stimulant, poly dA:dT, to the DC uptake receptor, DEC205 (DEC). We will show that inoculation of α-DEC-poly dA:dT fusion monoclonal antibodies (mAbs) is sufficient to induce rapid innate cytokine …
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Exploring the Roles of Heme Type and Histidine -Tyrosine Cross -Link in Heme-Copper Oxidases Using a Myoglobin Model
Novel oxygen reactivity is observed in our model protein (CuBMb) which reproduces the most essential components of HCO active site. In order to improve this artificial model to inhibit its side reaction with oxygen and channel it toward oxidase reactivity, more features were introduced into the …
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PNA-protein conjugates for nano-scale modeling of protein aggregates
Abstract Programmable assembly of proteins on molecular frameworks requires the development of facile and orthogonal chemical approaches and molecular scaffolds. In this research, the unique characteristics of PNA were applied to create controllable protein assemblies directed by precise PNA-DNA …
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Protein Semi-Synthesis in Vivo
<p>Incorporation of chemical probes into proteins is a powerful way to elucidate biological processes and to engineer novel function. This thesis describes the ligation of synthetic molecules to target proteins in an intracellular environment, and the use of semi-synthetic proteins for whole animal …
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Investigation of the mechanism of radical propagation in E. coli ribonucleotide reductase by site-specific incorporation of unnatural amino acids
… the best studied RNR to date and consists of two protein subunits, a2 and P2. a2 is the site of nucleotide reduction and 02 contains a diiron tyrosyl radical (Y122*) cofactor. Each turnover requires radical propagation from the Y122* in 32 to the active site of a2 over 35 A. The mechanism of this …
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Metalloprotein engineering with unnatural amino acids: application in functional heme-copper oxidase and azurin
Metalloproteins, proteins with metal ion cofactors, are estimated to make up more than a third of total proteome. They involve in key biological processes such as photosynthesis, respiration, and nitrogen fixation. The important roles and potential applications of metalloprotein urges the need for …
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Mechanistic Insights into the Stimulation of Dot1L-Mediated Methylation of Histone H3 by Semisynthetically Ubiquitylated Histone H2b
… of native uH2B. Two traceless orthogonal expressed protein ligation (EPL) reactions were used for this purpose, one employing a photolytically removable ligation auxiliary, and the other, a cysteine-mediated ligation followed by a desulfurization to restore the native sequence. …
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Exploring the Biophysical Properties of Domains Within Multi-Domain Proteins
… in understanding the biophysical properties of proteins using small modular domains such as SH3 domains. However, there is a surprising lack of knowledge regarding how these properties are affected when the domain is placed back within its full-length multi-domain protein. Using a combination of …
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Novel Interactions of the Hormone Leptin Revealed by PET Imaging in Rodents and Rhesus Macaques
… was introduced at the Cterminus of leptin using expressed protein ligation (EPL), which was subsequently derivitized with 18F-4-fluorobenzaldehyde in an aniline accelerated radiochemical oximation reaction. These probes were used for PET imaging in mice, rats, and in rhesus macaques. PET imaging …
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Development and application of chemical tools for investigating dynamic processes in cell migration
… offer insight into these important components of protein function. However, chemical tools, which function on a more rapid timescale and in localized regions of the cell, are capable of providing real-time information about protein activity. Herein, the development and application of chemical …
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Probing the roles of metal binding ligands in cupredoxins: incorporating nonproteinogenic amino acids into azurin and CuA Azurin
Item withdrawn by Mark Zulauf (zulauf@illinois.edu) on 2010-05-13T19:03:20Z Item was in collections: University of Illinois Theses & Dissertations (ID: 1) No. of bitstreams: 1 Clark_Kevin.pdf: 14900477 bytes, checksum: 901cc6f1c2f90be4a62e58094e6f0b6d (MD5)
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Semi-Synthesis of the Transcription Factor SMAD2 Containing Caging Groups and Phosphoaminoacid Analogues
<p>Post-translational modification (PTM) of a protein refers to any chemical change that occurs to the protein after its ribosomal synthesis. The seemingly endless number of PTMs can endow proteins with new functionalities that are not present in the unmodified proteins. In order to study the …
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Induktion von Sekundärstrukturen durch den Einbau von Alanyl-PNA in Peptide und Proteine
… eines Alanyl-PNA-Stranges in ein Peptid oder Protein und Zugabe des korrespondierenden Gegenstranges sollte zu einer lokalen Induktion eines β-Faltblattes führen und strukturelle Veränderungen im Gesamtpeptid hervorrufen. Es kann dann von einem molekularen Schalter gesprochen werden. Im Rahmen …
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Split Inteins: From Mechanistic Studies to Novel Protein Engineering Technologies
<p>Inteins are auto-processing protein domains that carry out a post-translational process known as protein splicing. This process is characterized by excision of the intein (intervening protein) domain from within a larger polypeptide sequence with concomitant ligation of the flanking extein ( …